The present invention relates to methods and compositions for the diagnosis, prevention, and treatment of cancer in mammals, and in particular, humans. More particularly, the present invention relates to methods and compositions for the diagnosis, prevention, and treatment of cancer through detection and modulation of the expression of TMF/ARA160 and Fer.
Cancer is one of the top killing diseases in the western world and vast amounts of effort and financial resources are being invested in developing novel therapeutic approaches. However, the need for reliable diagnostic tools, is a rate-limiting step in the successful application of a cancer therapy. This is best manifested by the fact that most of the currently known markers of cancers, are reliable at the level of only 30-50%. Thus the need for new markers that could be reliably used in the detection of a wide variety of cancers, exists Further, there is a generally accepted need for improved methods of cancer prevention and treatment, devoid of the well known side effects of current therapies. There is thus a widely recognized need for, and it would be highly advantageous to have, methods and compositions for the diagnosis of cancer that can distinguish the development of the malignant state and for methods and compositions for prevention and treatment of cancer.
A protein termed TMF or ARA60, which is present in a dormant form in normal mammalian cells has been recently identified (Garcia, J. A. et al. (1992) Proc. Natl. Acad. Sci. USA 89:9372-9376) [OMIM 601126, Locus ID 7110, GenBank L01042] Several functions have been attributed to TMF. It was initially identified as a DNA binding protein that preferentially binds to the TATA element in the human immunodeficiency 1 (HIV1) long terminal repeat (LTR) Thus, TMF/ARA160 was initially identified as a transcription factor that can suppress transcription of RNA Polymerase II genes by binding to their TATA box thus giving it the name TATA Element Regulatory Factor (TMF) (Garcia, J. A. et al. (1992) Proc. Natl. Acad. Sci. USA 89:9372-9376). Later, TMF was shown to function as a co-activator of nuclear receptors, particularly the androgen receptor (AR) (Hsiao, P. W. et al. (1999) J. Biol. Chem. 274:22373-22379), a fact that gained it the name androgen receptor coactivator 160 kDa, or ARA160 (Hsiao, P.-W. et al. (1999) J. Biol. Chem. 274:22373-22379).
TMF consists of 1093 amino acids with an apparent molecular mass of 160 kDa (Hsiao, P. W. at Chang, C. (1999) J. Biol. Chem. 274, 92373-29379; Garcia, J. A., Ou, S. H., Wu, F., Lusis, A. J., Sparkes, R. S. & Gaynor, R. B. (1992) Proc. Natl. Acad. Sci. USA 89, 9372-9376). The central and c-terminal parts of TMF/ARA160 contain coiled coil forming domains (cc) that could mediate the interaction of that protein with other cellular factors. Using a yeast two hybrid screening system (Schwartz, Y., Ben-Dor, I., Navon, A., Motro, B. & Nir, U. (1998) FEBS Lett 434, 339-345) it has been found that TMF/ARA160 interacts Edith Fer tyrosine kinases and modulates their activities. The Fer and AR binding domains in TMF/ARA160, overlap and both include cc forming sequences.
Fer (p94fer) is an evolutionarily conserved (Pawson, T., Letwin, K., Lee, T., Hao, Q.-L., Heisterkamp, N. & Groffen, J. (1989) Mol. Cell. Biol. 9, 5722-5725; Paulson, R., Jackson, J., Immergluck, K. & Bishop, J. M. (1997) Oncogene 14, 641-652) and ubiquitously expressed tyrosine kinase that resides mainly in the cytoplasm and nucleus of expressing cells (Letwin, K., Yee, S.-P. & Pawson, T. (1988) Oncogene 3, 621-627; Hao, Q.-L., Heisterkamp, N. & Groffen, J. (1989) Mol. Cell. Biol. 9, 1587-1593; Hao, Q.-L., Ferris, D. K., White, G., Heisterkamp, N. & Groffen, J. (1991) Mol. Cell. Biol. 11, 1180-1183; Kim, L. & Wong, T. W. (1998) J. Biol. Chem. 273, 23542-23548) [OMIM 176942, Locus ID 2241, GenBank J03358]. Fer vas not detected in mourn pert and T cell lines (Halachmy, S., Bern, O., Schreiber, L., Carmel, M., Sharabi, Y., Shoham, J. & Nir, U. (1997) Oncogene 14, 2871-2880).
In the cytoplasm, Fer associates with cell adhesion molecules (Kim, L. & Wong, T. W. (1998) J. Biol. Chem. 273, 23542-23548; Rosato, R., Veltmaat, J. M., Groffen, J. & Heisterkamp, N. (1998) Mol. Cell. Biol. 18, 5762-5770) and Stat3 (Priel-Halachmi, S., Ben-Dor, I., Shpungin, S., Tennenbaum, T., Molavani, H., Bachrach, M., Salzberg, S. & Nir, U. (2000) J. Biol Chem. 275, 28902-28910) and its kinase activity increases in growth factor stimulated cells (Kim, L and Wong, T. W (1995) Molecular & Cellular Biology). However, no direct role has been attributed to Fer in the establishment of adherens junctions or focal adhesions (Craig, A. W., Zimgibl, R., Williams, K. Cole, L. A & Greer, P. A. (2001) Mol. Cell Biol. 21, 603-613), nor was Fer found to be essential for growth factor dependent activation of Stat3. The function of Fer is redundant in the mouse, and mice devoid of a functional Fer are viable and fertile (Craig, A. W., Zimgibl, R., Williams, K., Cole, L A. & Greer, P. A. (2001) Mol. Cell Biol. 21, 603-613). However, the functioning of Fer was found to be pivotal for the proliferation of malignant cell lines (Allard, P., Zoubeidi, A, Nguyen, L. T., Tessier, S., Tanguay, S., Chevrette, M., Aprikian, A. & Chevalier, S. (2000) Mol. Cell Endocrinol. 159, 63-77; Orlovsky, K., Ben-Dor, I, Priel-Halachmi, S., Malovany, H. & Nir, U. (2000) Biochemistry 39, 11084-11091) Thus, Fer could be linked to the proliferation of mammalian cells.
A testis specific variant of Fer, termed p51ferT, is encoded by an alternatively spliced FER transcript (Fischman, K, Edman, J. C., Shackleford, G. M., Turner, J. A., Rutter, W. J. & Nir, U. (1990) Mol. Cell Biol 10, 146-153; Keshet, E., Itin, A, Fischman, K. & Nir, U. (1990) Mol. Cell. Biol. 10, 5021-5025). Fer and p51ferT share identical SH2 and kinase domains but they differ in their N-teminal tails (Hao, Q.-L., Heisterkamp, N. & Groffen, J. (1989) Mol. Cell. Biol. 9, 1587-1593; Fischman, K., Edman, J. C., Shackleford, G. M, Turner, J. A., Rutter, W. J. & Nir, U. (1990) Mol. Cell. Biol. 10, 146-153). p51ferT accumulates in late primary spermatocytes (Hazan, B., Bern, O., Carmel, M., Lejbkowicz, F., Goldstein, R. S. & Nir, U. (1993) Cell Growth Differ. 4, 443-449). However the role of that kinase in the spermatogenic process is also not understood (Craig, A. W., Zirngibl, R., Williams, K., Cole, L. A. & Greer, P. A. (2001) Mol. Cell Biol. 21, 603-613).
The activities attributed so far to TMF/ARA160 have not been linked to the Fer tyrosine kinase. Further, it has not heretofore been demonstrated that levels of expression of TMF/ARA160 can be measured or altered for diagnosis, prevention, or treatment of cancer. Specific modulation of expression of TMF/ARA160 and of Fer for prevention and treatment of cancer have not been developed Consequently there is an unmet need for agents and methods capable of effectively detecting the expression of TMF/ARA160 for the diagnosis of cancer and for modulation of expression of Fer and of TMF/ARA160 for prevention and treatment of cancer.